Immunization Against Pseudomonas aeruginosa by Modified Ad Vectors

描述(申请人提供)：本项目的目标是使用腺病毒(Ad)为基础的新型基因转移载体开发一种抗铜绿假单胞菌疫苗，该载体：编码铜绿假单胞菌外膜蛋白F(OprF)；已得到增强，在纤维中加入RGD序列，以增强通过？V？3.5整合素在体内向树突状细胞的输送；并将Epi8，一种保守的，高免疫原性的OprF表位，整合到六角外环。目的1重点是将这一策略用于基于Ad5的疫苗(Ad5cuOprF.RGD.Epi8)。我们预计它将引发高水平的抗P。血液和肺中的铜绿假单胞菌抗体，衣壳上的Epi8表位将允许加强免疫，即使存在对第一次免疫诱导的Ad的免疫。在完成毒理学测试并获得监管部门批准后，我们将在正常成年志愿者身上进行Ad5cuOprF.RGD.Epi8的临床试验。在目标2中，我们将进行AdC7cuOprF.RGD.Epi8，一种第二代抗P。与Ad5CuOprF.RGD.Epi8类似的铜绿假单胞菌疫苗，但基于非人类灵长类AdC7血清型，即使存在先前存在的抗Ad5免疫也应有效。该项目的具体目标是：目的1-测试Ad5cuOprF.RGD.Epi8是一种编码铜绿假单胞菌抗原OprF的人Ad5cuOprF.RGD.Epi8腺病毒，并用针对纤维中的RGD基序和OprF Epi8表位的树突状细胞进一步修饰的假设，是安全的，并将在健康人的血液和肺上皮衬里液中激发强大的抗假单胞菌调理抗体；目标2-检验有效抗P-P的假设。在AdC7cuOprF.RGD.Epi8抗Ad5免疫的背景下，可以实现铜绿假单胞菌免疫，AdC7cuOprF.RGD.Epi8是一种与Ad5cuOprF.RGD.Epi8相同的载体，但基于AdC7，一种非人类灵长类腺病毒，人类对其没有免疫力。铜绿假单胞菌呼吸道感染是囊性纤维化患者发病和死亡的主要原因。目前市场上尚无预防铜绿假单胞菌感染的疫苗，因此，成功的疫苗策略将对CF患者产生深远的影响。

DESCRIPTION (provided by applicant): The goal of this project is to develop an anti-Pseudomonas aeruginosa vaccine using a novel adenovirus (Ad)-based gene transfer vector that: codes for the P. aeruginosa outer membrane protein F (OprF); has been enhanced to incorporate an RGD sequence in the fiber to enhance delivery to dendritic cells in vivo via ?v?3.5 integrins; and incorporates Epi8, a conserved, highly immunogenic OprF epitope, into the hexon outer loop. Aim 1 focuses on using this strategy with an Ad5-based vaccine (Ad5cuOprF.RGD.Epi8). We expect that it will elicit high levels of anti-P. aeruginosa antibodies in blood and lung, and that the Epi8 epitope on the capsid will allow boosting, even in the presence of immunity against the Ad induced by the 1st immunization. After completing the toxicology testing, and gaining regulatory approval, we will carry out a clinical trial with Ad5cuOprF.RGD.Epi8 in normal adult volunteers. In aim 2, we will carry out studies with AdC7cuOprF.RGD.Epi8, a 2nd generation anti-P. aeruginosa vaccine that is similar to Ad5CuOprF.RGD.Epi8, but based on the non-human primate AdC7 serotype, a serotype that should be effective even in the presence of pre-existing anti-Ad5 immunity. The specific aims of the project are: Aim 1 - To test the hypothesis that administration of Ad5cuOprF.RGD.Epi8, a human Ad serotype 5 adenovirus that codes for the P. aeruginosa antigen OprF and is further modified with a dendritic cell targeting motif RGD in the fiber and the OprF Epi8 epitope in the hexon, is safe and will evoke robust anti-Pseudomonas opsonizing antibodies in blood and lung epithelial lining fluid of healthy individuals; Aim 2 - To examine the hypothesis that effective anti-P. aeruginosa immunity can be achieved in the context of anti-Ad5 immunity with AdC7cuOprF.RGD.Epi8, a vector identical to Ad5cuOprF.RGD.Epi8 but based on AdC7, a non-human primate adenovirus against which humans do not have immunity. Infections of the respiratory tract with Pseudomonas aeruginosa is the major cause of morbidity and mortality in individuals with cystic fibrosis. A marketed vaccine for prevention of infection with Pseudomonas aeruginosa is not available, therefore a successful vaccine strategy would have profound effects on individuals with CF.